Studies of Antibody Diversity and Genome Stability: Regulation of Activation-Indu

抗体多样性和基因组稳定性的研究：激活诱导的调节

• 批准号: 8002358
• 负责人: Yee Ling Wu
• 金额: $ 3.98万
• 依托单位: MEDICAL RES COUNCIL LAB OF MOLEC BIOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-11-24 至 2013-11-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8002358
• 关键词: Antibodies; Antibody Diversity; Biochemical; Cancer Etiology; Cell physiology; DNA; DNA Sequence; Disease; Enzymes; Fellowship; Generation of Antibody Diversity; Genes; Genetic Transcription; Genome Stability; Genomics; Goals; Hereditary Disease; Immune; Immunologics; Infection; Knowledge; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microbe; Molecular; Mutate; Mutation; Normal Cell; Organism; Physiological; Proteins; RNA Splicing; Regulation; Vertebrates; activation-induced cytidine deaminase; immune function; leukemia/lymphoma; microbial; next generation; novel therapeutics; public health relevance; research study

DESCRIPTION (provided by applicant): Activation-induced deaminase (AID) is an enzyme that mutates genes for antibodies in vertebrates. These AID-mediated mutations are necessary for an organism to generate specific antibodies to recognize and eradicate great varieties of microbial infections. However, if AID is not properly regulated, it can change DNA sequences of genes important for normal cell functions and cause diseases. Indeed, some lymphomas and leukemias are caused by abnormal activities of AID. How AID can change DNA sequences only in antibody genes but not in other genes is poorly understood. Recently, the catenin 2 like 1 protein (CTNNBL1) has been found to physically interact with AID. Little is known for the physiologic function of CTNNBL1. However, the interaction with CTNNBL1 is required for AID to change DNA sequences specifically in the antibody genes. In this study, a combination of high throughput approaches including next-generation sequencing and microarray analyses and targeted biochemical and immunologic analyses will be used to investigate how AID specifically mutate antibody genes, particularly what genomic regions are accessible to AID, both in the presence and in the absence of CTNNBL1. These experiments will also detect any alterations in transcription and splicing caused by the absence of CTNNBL1, and thereby shed lights on normal cellular functions of CTNNBL1. Mass spectrometry, mutagenic analyses and immunologic experiments will be performed to determine the molecular details of the interaction between AID and CTNNBL1 as well as to elucidate the mechanisms by which CTNNBL1 regulates the targeted activity of AID. It is anticipated that results from this study will be highly relevant for understanding the mechanisms underlying the generation of antibody diversity, and in the creation of new therapeutic strategies to minimize genomic disorders and cancers. PUBLIC HEALTH RELEVANCE: AID (activation-induced deaminase) is the DNA mutating enzyme responsible for changes of the antibody genes to generate different antibodies, but untimely or accidental actions of AID on cellular genes can cause cancers, immune-mediated or genetic diseases. The goal of this fellowship is to find out how AID is finely controlled to introduce mutations on genes for antibodies against microbes but not undesirable and harmful changes on genes of regular functions. Knowledge gained from this study will help understanding normal immune functions and facilitate the creation of new therapeutic strategies to minimize diseases such as cancers.

描述（由申请人提供）：活化诱导脱氨酶（AID）是一种使脊椎动物中抗体基因突变的酶。这些艾滋病介导的突变是生物体产生特异性抗体以识别和根除各种微生物感染所必需的。然而，如果AID没有得到适当的调节，它可以改变对正常细胞功能重要的基因的DNA序列并导致疾病。事实上，一些淋巴瘤和白血病是由AID的异常活动引起的。艾滋病如何只改变抗体基因的DNA序列，而不改变其他基因的DNA序列，目前还知之甚少。最近，已发现catenin 2 like 1蛋白（CTNNBL 1）与AID发生物理相互作用。CTNNBL 1的生理功能知之甚少。然而，与CTNNBL 1的相互作用是AID特异性改变抗体基因中的DNA序列所必需的。 在这项研究中，高通量方法的组合，包括下一代测序和微阵列分析和有针对性的生化和免疫学分析将被用来研究艾滋病如何特异性突变抗体基因，特别是哪些基因组区域是可访问的艾滋病，无论是在存在和不存在CTNNBL 1。这些实验还将检测由CTNNBL 1缺失引起的转录和剪接的任何改变，从而揭示CTNNBL 1的正常细胞功能。将进行质谱分析、致突变分析和免疫学实验，以确定AID和CTNNBL 1之间相互作用的分子细节，并阐明CTNNBL 1调节AID靶向活性的机制。 预计这项研究的结果将与理解抗体多样性产生的机制以及创建新的治疗策略以最大限度地减少基因组疾病和癌症高度相关。 公共卫生关系：AID（活化诱导脱氨酶）是DNA突变酶，负责抗体基因的变化以产生不同的抗体，但AID对细胞基因的不合时宜或意外作用可导致癌症、免疫介导或遗传疾病。这项研究的目标是找出艾滋病是如何被精细地控制，以引入基因突变的抗体对微生物，但不是不受欢迎的和有害的变化基因的正常功能。从这项研究中获得的知识将有助于了解正常的免疫功能，并促进创造新的治疗策略，以尽量减少癌症等疾病。